We are a group of 11 undergraduates representing Singapore as one of the 3 local teams for the International Genetically Engineered Machine 2018 (iGEM2018) competition.
The iGEM competition is an international synthetic biology competition which encourages multidisciplinary undergraduate students to come together, collaborate and address real-life problems through synthetic biology approaches. The competition is no simple feat as participants will be driven into a fast-paced working environment with only a short span of five months to present their experimental and modelling results. The competition will be held at Boston from 24 to 28 October 2018, where we will present our work to the international audience.
This year, our team is focused on creating a CRISPR-Cas13 system for Cytosine to Uracil RNA editing. Our project aims to extend the limited repertoire of proteins available for genetic editing by a proof of concept. This is to provide another tool in a toolbox for scientists to utilize. Currently, we're placed under the foundation track under iGEM.
We aim to construct a fusion protein using CRISPR-Cas13b as well as rAPOBEC-1. This fusion protein hosted in a mammalian plasmid backbone with gRNA will be transfected into HEK293T cells along with a dual reporter system. The dual reporter system is made up of GFP and mCherry connected by a T2A linker. To test the efficacy of our fusion protein, the start codon of GFP was mutated from AUG to ACG. Suppose that the fusion protein works, the fusion protein will be directed to the dual reporter and the rAPOBEC-1 domain will carry out deaminase activity to convert ACG to a functional start codon AUG. With a functional start codon, the GFP and mCherry (via T2A linker in the dual reporter) will be expressed, showing green and red fluorescence respectively. mCherry acts as a normalization factor to control for transfection and expression efficiency.
The CRISPR-Cas13 system utilises a sequence finder domain to seek out the specific mRNA strands, while the sequence editor modifies the Cytidine nucleotide to Uracil.
Apart from laboratory work, our team is actively involved in human practices. We believe that a constitutive link between ethics and research is one that will benefit the advancement of Science. Currently, we have conducted a series of interviews with experts in the field of genome editing. We will continue to bring out awareness within the community by posting regular YouTube updates, setting up information booths in public areas, as well as hosting workshops for local Secondary (Junior High) school students to pick up the basics of gene editing.
Experimental calibrations in progress.
Laboratory research constitutes the main bulk of our workload.
Genetic editing workshop conducted by our team member, Liyi at NUS High School.
Although a large proportion of the costs are covered by the institution and our benevolent sponsors, we are currently facing issues with the travelling expenses for the Boston trip. The trip may amount to a hefty SGD$19,800 for the entire team. Our funding goal of SGD$5,000 will suffice as 25% subsidy for the overall cost. Independent donations of any amount will help us ease the travel expenses and provide additional moral support to our team.
We hope that you can provide us with the support we need for this journey!
Do leave us a message if you have any queries!